Cytochrome P450 isoenzyme activities in cultured rat and mouse liver slices
Cytochrome P450 isoenzyme activities in cultured rat and mouse liver slices
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DOI：
10.1080/004982597240505
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发表时间：
1997-04-01
期刊：
影响因子：
1.8
通讯作者：
Yager, JD
中科院分区：
文献类型：
--
作者：
Gokhale, MS;Bunton, TE;Yager, JD
1.本研究的目的是测定几种细胞色素P450（CYP 450）同工酶的基础和诱导活性，并监测精确切割、培养的大鼠和小鼠肝切片中腺泡和肝细胞的形态.将切片在补充有胰岛素、转铁蛋白、硒、DMSO、地塞米松和表皮生长因子的Chee基本培养基中培养长达96小时。使用动态辊系统在37 ° C下在95%0（2）：5%CO2.3的气氛中孵育切片。肝切片的组织学显示，随着培养时间的延长，正常肝小叶结构得以维持。在培养的不同时间测定同工酶活性。在大鼠肝切片中，在72小时，CYP 1A 1/1X 2活性被β NF诱导4倍，被二恶英（TCDD）诱导37倍，而在小鼠肝切片中，1A 1/1A 2活性不能被β NF诱导，但被TCDD诱导19倍。在72 h时，在大鼠肝切片中未检测到CYP 2A 5（香豆素-7-羟化酶）活性，但在小鼠肝切片中，2A 5被β NF诱导2倍，被苯巴比妥（PB）诱导1倍，被TCDD诱导5倍。睾酮在特定位置的羟基化被用作各种P450亚型活性的指示。在0和72 h时向培养物中加入替吉奥，并在24和96 h时分别通过hplc分析测定代谢产物。根据种属、处理和培养时间，CYP 1A、2A、3A、2B和2C活性是令人满意的。PB在大鼠和小鼠肝切片中均高度诱导3A活性。这些结果表明，该培养系统可用于评估和比较啮齿类动物肝脏切片中的异生物质代谢。
1. The objective of this study was to determine the basal and inducible activities of several cytochrome P450 (CYP) isozymes and monitor the acinar and hepatocyte morphology in precision cut, cultured rat and mouse liver slices.2. The slices were cultured up to 96 h in Chee's essential medium supplemented with insulin, transferrin, selenium, DMSO, dexamethasone and epidermal growth factor. A dynamic roller system was used to incubate the slices at 37 degrees C in an atmosphere of 95% 0(2):5%, CO2.3. Histopathology of the liver slices revealed maintenance of normal hepatic lobular architecture with time in culture.4. CYP isozyme activities were measured at various times of culture. In rat liver slices, at 72 h, CYP1A1/1X2 activity was induced 4-fold by beta NF and 37-fold by dioxin (TCDD) whereas in mouse liver slices, 1A1/1A2 activity was not inducible by beta NF but was induced 19-fold by TCDD. At 72 h, CYP2A5 (coumarin-7-hydroxylase) activity was not detected in rat liver slices but in mouse liver slices, 2A5 was induced 2-fold by beta NF, Ii-fold by phenobarbital (PB) and S-fold by TCDD.5. Hydroxylation of testosterone at specific positions was used as an indication of the activities of various P450 isoforms. Testosterone was added to the cultures at 0 and 72 h and the metabolites were measured at 24 and 96 h respectively by hplc analysis. Depending upon the species, the treatment and the time in culture, CYP1A, 2A, 3A, 2B and 2C activities were delectable. 3A activity was highly induced by PB in both rat and mouse liver slices. These results demonstrate that this culture system can be used to assess and compare xenobiotic metabolism in liver slices from rodent species.